Acta Agrestia Sinica ›› 2021, Vol. 29 ›› Issue (11): 2591-2599.DOI: 10.11733/j.issn.1007-0435.2021.11.026

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Establishment of Hairy Root Transformation System of Melilotus albus Induced by Agrobacterium rhizogenes

WANG Sheng-sheng, DUAN Zhen, ZHANG Ji-yu   

  1. State Key Laboratory of Grassland Agro-Ecosystems, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, Gansu Province 730020, China
  • Received:2021-06-07 Revised:2021-08-16 Online:2021-11-15 Published:2021-12-06

发根农杆菌介导的白花草木樨毛状根转化体系的建立

王升升, 段珍, 张吉宇   

  1. 兰州大学草地农业生态系统国家重点实验室, 兰州大学草地农业科技学院, 甘肃 兰州 730020
  • 通讯作者: 张吉宇,E-mail:zhangjy@lzu.edu.cn
  • 作者简介:王升升(1996-),女,汉族,甘肃天水人,硕士研究生,主要从事草类作物遗传育种研究,E-mail:wangshsh19@lzu.edu.cn
  • 基金资助:
    国家自然科学基金-国际(地区)合作与交流项目(NSFC-CGIAR)(32061143035)和甘肃省科技重大专项(19ZD2NA002-2)资助

Abstract: Melilotus albus is one of the most common cultivated species of M. spp,which has high resistance and nitrogen fixation ability. In this study,we established an efficient hairy root transformation system induced by Agrobacterium rhizogenes in M. albus,and the nodulation gene MaROP6 was used as the target gene. The results showed that hairy roots were obtained within two weeks with a rate of 78%,and the positive transgenic hairy roots of MaROP6 gene reached 70.8%. The qRT-PCR results showed that the average relative expression of MaROP6 in transgenic hairy roots was 36.87 times higher than that in the control. The establishment of hairy root transformation system laid a foundation for identifying the gene function of M. albus.

Key words: Melilotus albus, Agrobacterium rhizogenes, Hairy root transformation, Gene expression

摘要: 白花草木樨(Melilotus albus Medic.ex Desr.)是草木樨最常见的栽培用种之一,具有较强的抗逆性和固氮能力。为了开发一套高效的草木樨转化体系,本研究以白花草木樨为材料,以结瘤基因MaROP6为目的基因,建立了发根农杆菌(Agrobacterium rhizogenes)介导的高效白花草木樨毛状根转化体系。结果表明:利用该方法可以在两周左右获得毛状根,其发生率可达78%,MaROP6基因的毛状根转化率可达70.8%。qRT-PCR鉴定结果表明,MaROP6转基因毛状根的平均相对表达量为对照的36.87倍。毛状根转化体系的建立为白花草木樨基因功能的验证奠定了基础。

关键词: 白花草木樨, 发根农杆菌, 毛状根转化, 基因表达

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